Separation, Sizing, and Quantitation of Engineered Nanoparticles in an Organism Model Using Inductively Coupled Plasma Mass Spectrometry and Image Analysis.
Separation, Sizing, and Quantitation of Engineered Nanoparticles in an Organism Model Using Inductively Coupled Plasma Mass Spectrometry and Image Analysis.
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DOI：
10.1021/acsnano.6b06582
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发表时间：
2017-01-24
期刊：
影响因子：
17.1
通讯作者：
Nelson BC
中科院分区：
文献类型：
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作者：
Johnson ME;Hanna SK;Montoro Bustos AR;Sims CM;Elliott LC;Lingayat A;Johnston AC;Nikoobakht B;Elliott JT;Holbrook RD;Scott KC;Murphy KE;Petersen EJ;Yu LL;Nelson BC
消费产品中越来越多地使用工程纳米颗粒(ENPs)，而且相对缺乏对其相关危害的了解，因此有必要进一步研究ENPs与环境和生物系统之间的相互作用。对于评估口服摄入的ENPs的环境研究，确保准确定量摄入的ENPs的一个关键步骤是有效地将有机体与ENPs分离，这些ENPs要么非特异性地吸附在有机体上，要么在暴露后悬浮在分散体中。在这里，我们测量了自由生活的线虫秀丽线虫对30 nm和60 nm金纳米颗粒(AuNPs)的摄取，使用蔗糖密度梯度离心法去除未摄入的AuNPs。采用常规电感耦合等离子体质谱(ICP-MS)和单粒子(SP)电感耦合等离子体质谱(SP-MS)分别测量了摄入的AuNPs的总质量和粒度分布。扫描电子显微镜/能量色散X射线能谱(SEM/EDS)成像证实，传统的线虫清洗方法在去除暴露后多余的悬浮和/或吸附的AuNPs方面无效。在非暴露和暴露于Au的线虫的酸性消化后，定量测定总的Au吸收，而碱性消化程序被优化，以释放摄入的AuNPs用于SPICPMS表征。通过高压冷冻/冷冻替代树脂制备和大面积扫描电子显微镜成像，测定了线虫摄取AuNPs的粒度分布和颗粒数浓度，相应地证实了线虫摄取AuNPs。在多细胞生物体中分离和体内量化ENP的方法将有助于对ENP在环境中的摄取、生物转化和危害评估进行强有力的研究。
The increased use of engineered nanoparticles (ENPs) in consumer products and the relative lack of understanding of their associated hazards necessitate further investigation of the interactions between ENPs and environmental and biological systems. For environmental studies assessing uptake of orally ingested ENPs, a key step in ensuring accurate quantification of ingested ENPs is efficient separation of the organism from ENPs that are either nonspecifically adsorbed to the organism and/or suspended in the dispersion following exposure. Here, we measure the uptake of 30 nm and 60 nm gold nanoparticles (AuNPs) by the free-living nematode, Caenorhabditis elegans, using a sucrose density gradient centrifugation protocol to remove non-ingested AuNPs. Both conventional inductively coupled plasma-mass spectrometry (ICP-MS) and single particle (sp) ICP-MS are utilized to measure the total mass and size distribution, respectively, of ingested AuNPs. Scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDS) imaging confirmed that traditional nematode washing procedures were ineffective at removing excess suspended and/or adsorbed AuNPs after exposure. Quantification of total Au uptake was performed following acidic digestion of non-exposed and Au-exposed nematodes, while an alkaline digestion procedure was optimized for the liberation of ingested AuNPs for spICP-MS characterization. Size distributions and particle number concentrations were determined for AuNPs ingested by nematodes with corresponding confirmation of nematode uptake via high-pressure freezing/freeze substitution resin preparation and large-area SEM imaging. Methods for the separation and in vivo quantification of ENPs in multicellular organisms will facilitate robust studies of ENP uptake, biotransformation, and hazard assessment in the environment.